Background/Objectives: Adipose tissue (AT) plays significant roles in energy storage, metabolite signaling, and immunomodulation. The understanding of its underlying mechanisms of dysregulation can provide insight into complex disease processes through analysis with histology, flow cytometry, metabolomics, and proteomics. Tissue sampling in the clinical setting has largely shifted towards minimally invasive approaches to improve factors such as patient satisfaction, post-operative recovery, and procedure length. In contrast, preclinical animal models continue to rely on more invasive methods until refined, minimally invasive techniques are developed and systematically assessed. To improve animal welfare and enhance clinical translatability, there is a critical need to reverse translate these approaches into animal models. Methods: Our study evaluated the feasibility and performance of a commercially available vacuum-assisted biopsy (VAB) device for AT sampling in a preclinical nonhuman primate (NHP) model. Six rhesus NHPs successfully underwent three serial AT biopsies with a VAB device (n = 18). Results: All animals recovered without any serious or unexpected adverse events. The amount of adipose tissue collected per biopsy (0.5–2.7 g) was proportional to the number of individual tracks. Isolation of the stromal vascular fraction (SVF) from a subset of samples (n = 6) yielded 0.41 ± 0.12 × 10⁶ cells/g of tissue. Conclusions: The minimally invasive VAB technique is a safe and reliable method of AT collection in NHPs. This feasibility study demonstrated adequate volumes of tissue cores that are suitable for typical, downstream research applications including immunologic studies and pathology, while improving animal welfare.